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1.
Front Plant Sci ; 15: 1378079, 2024.
Artigo em Inglês | MEDLINE | ID: mdl-38947947

RESUMO

Introduction: The Salkowski reagent method is a colorimetric technique used to determine auxin production, specifically as indole-3-acetic acid (IAA). It was developed to determine indoles rapidly; however, it does not follow Beer's law at high concentrations of IAA. Thus, there could be an overestimation of IAA with the Salkowski technique due to the detection of other indole compounds. Methods: This study aims to compare the Salkowski colorimetric method versus a chromatographic method to evidence the imprecision or overestimation obtained when auxins, such as indole-acetic acid (IAA), are determined as traits from promoting growth plant bacteria (PGPB), using ten different strains from three different isolation sources. The analysis used the same bacterial culture to compare the Salkowski colorimetric and chromatographic results. Each bacterium was cultivated in the modified TSA without or with tryptophan for 96 h. The same supernatant culture was used in both methods: Salkowski reagent and ultra-performance liquid chromatography coupled with a Mass Spectrometer (LC-MS/MS). Results: The first method indicated 5.4 to 27.4 mg L-1 without tryptophan in ten evaluated strains. When tryptophan was used as an inductor of auxin production, an increase was observed with an interval from 4.4 to 160 mg L-1. The principal auxin produced by all strains was IAA from that evaluated by the LC-MS/MS method, with significantly higher concentration with tryptophan addition than without. Strains belonging to the Kocuria genus were highlighted by high IAA production. The indole-3-propionic acid (IPA) was detected in all the bacterial cultures without tryptophan and only in K. turfanensis As05 with tryptophan, while it was not detected in other strains. In addition, indole-3-butyric acid (IBA) was detected at trace levels (13-16 µg L-1). Conclusions: The Salkowski reagent overestimates the IAA concentration with an interval of 41-1042 folds without tryptophan and 7-16330 folds with tryptophan as inductor. In future works, it will be necessary to determine IAA or other auxins using more suitable sensitive techniques and methodologies.

2.
Plants (Basel) ; 12(14)2023 Jul 08.
Artigo em Inglês | MEDLINE | ID: mdl-37514205

RESUMO

The search for sustainable agriculture has increased interest in using endophytic bacteria to reduce fertilizer use and increase stress resilience. Stress-adapted plants are a potential source of these bacteria. Some species of these plants have not yet been evaluated for this, such as pangolão grass, from which we considered endophytic bacteria as potential plant growth promoters. Bacteria from the root, colm, leaves, and rhizospheric soil were isolated, and 132 strains were evaluated for their in vitro biological nitrogen fixation, IAA and siderophores production, and phosphate solubilization. Each mechanism was also assessed under low N availability, water stress, and low-solubility Fe and P sources in maize greenhouse experiments. All strains synthesized IAA; 63 grew on N-free media, 114 synthesized siderophores, and 46 solubilized P, while 19 presented all four mechanisms. Overall, these strains had better performance than commercial inoculant in all experiments. Still, in vitro responses were not good predictors of in vivo effects, which indicates that the former should not be used for strain selection, since this could lead to not testing strains with good plant growth promotion potential. Their heterologous growth promotion in maize reinforces the potential of stress-adapted plant species as potential sources of strains for inoculants.

3.
Microbiol Res ; 266: 127218, 2023 Jan.
Artigo em Inglês | MEDLINE | ID: mdl-36242861

RESUMO

The bacterial biosynthesis of indole-3-acetic acid (IAA) is often related to the beneficial effects of plant growth-promoting rhizobacteria (PGPR) on plant development. In PGPR belonging to the Bacillus genus, the synthesis of IAA may occur through different metabolic pathways that are still poorly understood. B. thuringiensis (Bt) is well known for its insecticidal properties; however, its beneficial features are not limited to pest control. Our group has been studed the beneficial effects of Bt strain RZ2MS9 as growth promoter in a range of plant crops, including soybean, tomato, and maize. We recently demonstrated that bacterial IAA biosynthesis plays an important role in the ability of RZ2MS9 to benefit plant development. However, the molecular involved mechanisms in the IAA biosynthesis by this bacterium in the beneficial interaction with plants remain unclear. Here, we investigated the genetic basis of IAA biosynthesis by RZ2MS9. We knocked out the ipdC gene, involved in IAA biosynthesis via the tryptophan-dependent IPyA pathway, using the CRISPR-Cas9 system. Our results showed that, by disrupting the IPyA pathway, the amount of IAA synthesized by the mutant RZ2MS9 (ΔipdC) in the presence of tryptophan drops 57%. The gene knockout did not affect the bacterial growth, but it did affect its ability to colonize maize. Moreover, deactivating the ipdC gene in RZ2MS9 significantly reduces its ability to promote maize growth. ΔipdC performed worse than RZ2MS9 in almost all evaluated plant parameters, including total root length, projected root area, lateral roots, aerial part dry matter, and germination speed index. Therefore, we demonstrated that tryptophan-dependent IAA biosynthesis via the IPyA pathway by RZ2MS9 is strongly influenced by the ipdC gene. Furthermore, IAA biosynthesis by RZ2MS9 is a major mechanism used by this PGPR to promote maize growth.


Assuntos
Bacillus thuringiensis , Zea mays , Zea mays/genética , Zea mays/metabolismo , Reguladores de Crescimento de Plantas/metabolismo , Bacillus thuringiensis/genética , Bacillus thuringiensis/metabolismo , Triptofano/metabolismo , Técnicas de Inativação de Genes , Sistemas CRISPR-Cas , Ácidos Indolacéticos/metabolismo
4.
Mycology ; 13(4): 257-270, 2022.
Artigo em Inglês | MEDLINE | ID: mdl-36405335

RESUMO

The protective and growth-promoting activities of Colletrotrichum and Diaporthe endophytes on tomato plants (Lycopersicon esculentum Mill.) are underexplored. We screened 40 endophytic fungi associated with Mexican shrimp plant (Justicia brandegeana) using an in vitro dual culture assay for Fusarium oxysporum, one of the most important phytopathogens of tomato plants. The three best antagonists, Colletotrichum siamense (JB224.g1), C. siamense (JB252.g1), and Diaporthe masirevicii (JB270), were identified based on multilocus sequence analysis. They were assessed in vitro for their inhibition of F. oxysporum and phosphate solubilisation capacity, and for the production of indole acetic acid. Greenhouse experiments verified the growth-promoting effects of these endophytes and the suppression of F. oxysporum symptoms in tomato plants.   Under greenhouse conditions, the JB252.g1 and JB270 isolates showed positive results for seedling emergence speed. The radicular system depth of plants inoculated with JB270 was greater than that in uninoculated plants (27.21 vs 21.95 cm). The soil plant analysis development chlorophyll metre (SPAD) index showed statistically significant results, especially for the endophyte JB224.g1 (36.99) compared to the control plants (30.90) and plants infected solely with F. oxysporum (33.64).

5.
Plants (Basel) ; 11(11)2022 May 31.
Artigo em Inglês | MEDLINE | ID: mdl-35684245

RESUMO

The capacity for apomixis in Paspalum notatum is controlled by a single-dominant genomic region, which shows strong synteny to a portion of rice chromosome 12 long arm. The locus LOC_Os12g40890, encoding the Auxin/Indole-3-Acetic Acid (Aux/IAA) family member OsIAA30, is located in this rice genomic segment. The objectives of this work were to identify transcripts coding for Aux/IAA proteins expressed in reproductive tissues of P. notatum, detect the OsIAA30 putative ortholog and analyze its temporal and spatial expression pattern in reproductive organs of sexual and apomictic plants. Thirty-three transcripts coding for AUX/IAA proteins were identified. Predicted protein alignment and phylogenetic analysis detected a highly similar sequence to OsIAA30 (named as PnIAA30) present in both sexual and apomictic samples. The expression assays of PnIAA30 showed a significant down-regulation in apomictic spikelets compared to sexual ones at the stages of anthesis and post-anthesis, representation levels negatively correlated with apospory expressivity and different localizations in sexual and apomictic ovules. Several PnIAA30 predicted interactors also appeared differentially regulated in the sexual and apomictic floral transcriptomes. Our results showed that an auxin-response repressor similar to OsIAA30 is down-regulated in apomictic spikelets of P. notatum and suggests a contrasting regulation of auxin signaling during sexual and asexual seed formation.

6.
Front Microbiol ; 13: 1062414, 2022.
Artigo em Inglês | MEDLINE | ID: mdl-36741893

RESUMO

Introduction: Plants interact with plant growth-promoting bacteria (PGPB), especially under stress condition in natural and agricultural systems. Although a potentially beneficial microbiome has been found associated to plants from alpine systems, this plant- PGPB interaction has been scarcely studied. Nevados de Chillán Complex hold one of the southernmost xerophytic formations in Chile. Plant species living there have to cope with drought and extreme temperatures during the growing season period, microclimatic conditions that become harsher on equatorial than polar slopes, and where the interaction with PGPB could be key for plant survival. Our goal was to study the abundance and activity of different PGPB associated to two abundant plant species of Andean xerophytic formations on contrasting slopes. Methods: Twenty individuals of Berberis empetrifolia and Azorella prolifera shrubs were selected growing on a north and south slope nearby Las Fumarolas, at 2,050 m elevation. On each slope, microclimate based on temperature and moisture conditions were monitored throughout the growing period (oct. - apr.). Chemical properties of the soil under plant species canopies were also characterized. Bacterial abundance was measured as Log CFU g-1 from soil samples collected from each individual and slope. Then, the most abundant bacterial colonies were selected, and different hormonal (indoleacetic acid) and enzymatic (nitrogenase, phosphatase, ACC-deaminase) mechanisms that promote plant growth were assessed and measured. Results and Discussion: Extreme temperatures were observed in the north facing slope, recording the hottest days (41 vs. 36°C) and coldest nights (-9.9 vs. 6.6°C). Moreover, air and soil moisture were lower on north than on south slope, especially late in the growing season. We found that bacterial abundance was higher in soils on north than on south slope but only under B. empetrifolia canopy. Moreover, the activity of plant growth-promoting mechanisms varied between slopes, being on average higher on north than on south slope, but with plant species-dependent trends. Our work showed how the environmental heterogeneity at microscale in alpine systems (slope and plant species identity) underlies variations in the abundance and plant growth promoting activity of the microorganisms present under the plant canopy of the Andean xerophytic formations and highlight the importance of PGPB from harsh systems as biotechnological tools for restoration.

7.
World J Microbiol Biotechnol ; 37(10): 164, 2021 Aug 30.
Artigo em Inglês | MEDLINE | ID: mdl-34458956

RESUMO

Eucalyptus is the main species for the forestry industry in Brazil. Biotechnology and, more recently, gene editing offer significant opportunities for rapid improvements in Eucalyptus breeding programs. However, the recalcitrance of Eucalyptus species to in vitro culture is also a major limitation for commercial deployment of biotechnology techniques in Eucalyptus improvement. We evaluated various clones of Eucalyptus urophylla for their in vitro regeneration potential identified a clone, BRS07-01, with considerably higher regeneration rate (85%) in organogenesis, and significantly higher than most works described in literature. Endophytic bacteria are widely reported to improve in vitro plant growth and development. Hence, we believe that inclusion of endophytic plant growth promoting bacteria enhanced was responsible for the improved plantlets growth and development of this clone under in vitro culture. Metagenomic analysis was performed to isolate and characterize the prominent endophytic bacteria on BRS07-01 leaf tissue in vitro micro-cultures, and evaluate their impact on plant growth promotion. The analysis revealed the presence of the phyla Firmicutes (35%), Proteobacteria (30%) and much smaller quantities of Actinobacteria, Bacteroidetes, Gemmatimonadetes, Crenarchaeota, Euryarchaeota and Acidobacteria. Of the thirty endophytic bacterial strains isolated, eleven produced indole-3-acetic acid. Two of the isolates were identified as Enterobacter sp. and Paenibacillus polymyxa, which are nitrogen-fixing and capable of phosphate and produce ammonium. These isolates also showed similar positive effects on the germination of common beans (Phaseolus spp.). The isolates will now be tested as a growth promoter in Eucalyptus in vitro cultures. Graphical abstract for the methodology using cultivation independent and dependent methodologies to investigate the endophytic bacteria community from in vitro Eucalyptus urophylla BRS07-01.


Assuntos
Bactérias/isolamento & purificação , Endófitos/isolamento & purificação , Eucalyptus/microbiologia , Bactérias/classificação , Bactérias/genética , Bactérias/metabolismo , Brasil , DNA Bacteriano/genética , DNA Ribossômico/genética , Endófitos/classificação , Endófitos/genética , Endófitos/metabolismo , Eucalyptus/crescimento & desenvolvimento , Ácidos Indolacéticos/metabolismo , Metagenômica , Filogenia , Folhas de Planta/crescimento & desenvolvimento , Folhas de Planta/microbiologia , RNA Ribossômico 16S/genética
8.
Ecotoxicol Environ Saf ; 218: 112273, 2021 Apr 30.
Artigo em Inglês | MEDLINE | ID: mdl-33940441

RESUMO

The aim of the study was to characterize halotolerant bacteria and to evaluate their plant growth promotion potential on chia and quinoa seedlings under saline stress. Isolated microorganisms were evaluated for nitrogen fixation, phosphate solubilization, and production of siderophores and indole acetic acid. Three strains and two consortia were selected: Halomonas sp. (SFS), Micrococcus luteus (SA211), Bacillus sp. (HX11), C1 (SA211 + SFS), and C2 (SA211 + HX11). In vitro assays using water agar and half-strength Murashige-Skoog plates showed that an increase in salinity led to an increased seedlings mortality and a decrease in germination (lower than 40%), in total length (varying between 16% and 87% decreases), root length (from 60% to 92% lesser length) and dry weight (from 7% to 86% lower weight). Also, the relative growth index (RGI) decreased for both crops in most treatments, except those with HX11 and C2. These treatments had the highest growth parameters and RGI values in presence of high salinity in chia (50 and 100 mmol/L NaCl) and quinoa (200 and 400 mmol/L NaCl). SA211, the highest producer of indole acetic acid, showed a detrimental effect and anomalous phenotype on plants. Our results suggest that Bacillus sp. HX11, with multiple plant growth promotion traits and tolerance to saline stress, has a great potential as a bioinoculant in saline conditions and could be used as a biofertilizer for crop production.

9.
World J Microbiol Biotechnol ; 37(1): 9, 2021 Jan 04.
Artigo em Inglês | MEDLINE | ID: mdl-33392828

RESUMO

Current worldwide challenges are to increase the food production and decrease the environmental contamination by industrial emissions. For this, bacteria can produce plant growth promoter phytohormones and mediate the bioremediation of sewage by heavy metals removal. We developed a Rational Design of Immobilized Derivatives (RDID) strategy, applicable for protein, spore and cell immobilization and implemented in the RDID1.0 software. In this work, we propose new algorithms to optimize the theoretical maximal quantity of cells to immobilize (tMQCell) on solid supports, implemented in the RDIDCell software. The main modifications to the preexisting algorithms are related to the sphere packing theory and exclusive immobilization on the support surface. We experimentally validated the new tMQCell parameter by electrostatic immobilization of ten microbial strains on AMBERJET® 4200 Cl- porous solid support. All predicted tMQCell match the practical maximal quantity of cells to immobilize with a 10% confidence. The values predicted by the RDIDCell software are more accurate than the values predicted by the RDID1.0 software. 3-indolacetic acid (IAA) production by one bacterial immobilized derivative was higher (~ 2.6 µg IAA-like indoles/108 cells) than that of the cell suspension (1.5 µg IAA-like indoles/108 cells), and higher than the tryptophan amount added as indole precursor. Another bacterial immobilized derivative was more active (22 µg Cr(III)/108 cells) than the resuspended cells (14.5 µg Cr(III)/108 cells) in bioconversion of Cr(VI) to Cr(III). Optimized RDID strategy can be used to synthesize bacterial immobilized derivatives with useful biotechnological applications.


Assuntos
Biodegradação Ambiental , Células Imobilizadas/metabolismo , Biologia Computacional/métodos , Algoritmos , Bactérias/metabolismo , Biomassa , Poluentes Ambientais , Metais Pesados/metabolismo , Software , Eletricidade Estática
10.
Acta Sci. Biol. Sci. ; 43: e51737, 2021. tab, ilus
Artigo em Inglês | VETINDEX | ID: vti-764583

RESUMO

Endophytes are growth-promoting agents capable of synthesizing phytohormones, uptaking nutrients, and controlling pathogens. There is a strong potential to exploit them in the agriculture field like biofertilizers and biocontrol agents. In this work, we aimed to evaluate endophytic fungi isolated from Pachystachys lutea for their potential to solubilize phosphate, synthesise indole acetic acid (IAA), antagonize phytopathogens, and promote plant growth under greenhouse conditions. The phosphate solubilization efficiency was assessed on Pikovskayas agar medium. For analysis of IAA production, mycelia plugs of endophytes were cultured in Potato Dextrose Broth medium supplemented with L-tryptophan, with Salkowski Reagent, and the absorbance of the culture was measured. The antagonism evaluation of strain Alternaria sp. PL75 against phytopathogens was performed using the paired-culture technique. The promotion of plant growth provided by Alternaria sp. PL75 was evaluated in tomato plants. All strains evaluated were able to solubilize phosphate; however, the strain Alternaria sp. PL75 was the most effective (4.29). Two strains, Nemania sp. PL27 and Alternaria sp. PL75, produced 1.86 and 1.73 & 956;g mL-1 of IAA, respectively. In the antagonism assay, the endophyte Alternaria sp. PL75 and its fungal extract showed the best results against the pathogen Moniliophthora perniciosa. The greenhouse experiment result showed the endophyte Alternaria sp. PL75 increased the plantlets emergency speed index and the percentage of germination from 60 to 81.63%. It was also observed a statistical significance in the shoot length of the treated plants with the endophyte suspension (55.38 cm) compared to the control (41.67 cm).(AU)


Assuntos
Lamiales/crescimento & desenvolvimento , Endófitos , Ácido Acético/análise , Fosfatos
11.
Acta amaz. ; 51(1): 10-19, mar. 2021. tab, graf, ilus
Artigo em Inglês | VETINDEX | ID: vti-30253

RESUMO

Despite advances in the identification and characterization of endophytic bacteria in various plant species worldwide, little is known about such microorganisms in plants from the Amazon region. Previous studies reported that Piper tuberculatum endophytic Pseudomonas (isolates Pt12 and Pt13, identified as Pseudomonas putida and Pseudomonas sp., respectively) were able to inhibit the in vitro growth of Fusarium solani f. sp. piperis, which causes root rot in black pepper (Piper nigrum), and that Pt13 promoted the growth of P. nigrum. Therefore, the aim here was to characterize these bacteria regarding their ability to produce plant growth-promoting substances [siderophores, indol acetic acid (IAA) and soluble phosphate]. Chrome azurol S assays were performed for the detection of siderophores. For qualitative and quantitative assays of IAA production and phosphate solubilization, Salkowski´s reagent and NBRIP medium with molybdenum blue reagent, respectively, were used. Results revealed that Pt12 and Pt13 were able to synthesize IAA, mainly under a high concentration of L-tryptophan, indicating that they are IAA-producing bacteria, probably through a tryptophan-dependent biosynthesis pathway. The presence of P. nigrum extract positively influenced the IAA production by Pt12 and Pt13, with highest values of 125 and 90 µg mL-1, respectively. In addition, Pt12 was positive for the production of siderophores and produced 56.56 µg mL-1 of soluble phosphate. In contrast, Pt13 showed no ability to produce siderophores or to solubilize phosphate. Besides their potential in controlling plant diseases, Pt12 and Pt13 have potential as biofertilizers, favoring sustainable agriculture.(AU)


Apesar dos avanços na identificação e caracterização de bactérias endofíticas em espécies vegetais em todo o mundo, pouco se sabe sobre esses microrganismos em plantas da região amazônica. Estudos anteriores mostraram que Pseudomonas de Piper tuberculatum (isolados Pt12 e Pt13, identificados como Pseudomonas putida e Pseudomonas sp., respectivamente) são capazes de inibir Fusarium solani f. sp. piperis, que causa a podridão das raízes da pimenteira-do-reino (Piper nigrum), e que Pt13 promoveu o crescimento de P. nigrum. Portanto, o objetivo do presente trabalho foi caracterizar essas bactérias quanto à capacidade de produzir substâncias potencialmente bioestimulantes para o crescimento vegetal [sideróforos, ácido indol acético (AIA) e fosfato solúvel]. Ensaios de Cromo Azurol S foram realizados para detecção de sideróforos. Para os ensaios qualitativos e quantitativos de produção de AIA e solubilização de fosfato, foram utilizados o reagente de Salkowski e o meio NBRIP com azul de molibdênio, respectivamente. Os resultados revelaram que Pt12 e Pt13 sintetizaram AIA, principalmente sob alta concentração de L-triptofano, indicando que provavelmente utilizam uma via de biossíntese dependente deste aminoácido. A presença do extrato de P. nigrum influenciou positivamente a produção de AIA por Pt12 e Pt13, com valores máximos de 125 e 90 µg mL-1, respectivamente. Além disso, Pt12 foi positiva para produção de sideróforos e produziu 56.56 µg.mL-1 de fosfato solúvel. Em contraste, Pt13 não produziu sideróforos, nem solubilizou fosfato. Além do potencial de controle de doenças de plantas, Pt12 e Pt13 têm potencial como biofertilizantes, favorecendo a agricultura sustentável.(AU)


Assuntos
Piper nigrum/microbiologia , Endófitos/química , Pseudomonas/classificação , Sideróforos , Ácidos Indolacéticos
12.
Acta sci., Biol. sci ; Acta sci., Biol. sci;43: e51737, 2021. tab, ilus
Artigo em Inglês | LILACS, VETINDEX | ID: biblio-1460974

RESUMO

Endophytes are growth-promoting agents capable of synthesizing phytohormones, uptaking nutrients, and controlling pathogens. There is a strong potential to exploit them in the agriculture field like biofertilizers and biocontrol agents. In this work, we aimed to evaluate endophytic fungi isolated from Pachystachys lutea for their potential to solubilize phosphate, synthesise indole acetic acid (IAA), antagonize phytopathogens, and promote plant growth under greenhouse conditions. The phosphate solubilization efficiency was assessed on Pikovskaya’s agar medium. For analysis of IAA production, mycelia plugs of endophytes were cultured in Potato Dextrose Broth medium supplemented with L-tryptophan, with Salkowski Reagent, and the absorbance of the culture was measured. The antagonism evaluation of strain Alternaria sp. PL75 against phytopathogens was performed using the paired-culture technique. The promotion of plant growth provided by Alternaria sp. PL75 was evaluated in tomato plants. All strains evaluated were able to solubilize phosphate; however, the strain Alternaria sp. PL75 was the most effective (4.29). Two strains, Nemania sp. PL27 and Alternaria sp. PL75, produced 1.86 and 1.73 & 956;g mL-1 of IAA, respectively. In the antagonism assay, the endophyte Alternaria sp. PL75 and its fungal extract showed the best results against the pathogen Moniliophthora perniciosa. The greenhouse experiment result showed the endophyte Alternaria sp. PL75 increased the plantlets emergency speed index and the percentage of germination from 60 to 81.63%. It was also observed a statistical significance in the shoot length of the treated plants with the endophyte suspension (55.38 cm) compared to the control (41.67 cm).


Assuntos
Endófitos , Fosfatos , Lamiales/crescimento & desenvolvimento , Ácido Acético/análise
13.
Saudi J Biol Sci ; 27(8): 1913-1922, 2020 Aug.
Artigo em Inglês | MEDLINE | ID: mdl-32714014

RESUMO

Certain soil bacteria produce beneficial effects on the growth and health of plants; hence, their use is steadily increasing. Five strains of Bacillus with plant growth-promoting potential were selected in this study, which produced indole-3-acetic acid levels below 50 µg.mL-1. On the other hand, while only strains M8 and M15 dissolved phosphorus, the latter was the only strain that did not produce siderophores. Only strains M8 and M16 significantly inhibited the in vitro growth of Botrytis cinerea and Fusarium solani phytopathogens, whose inhibition ranges fluctuated between 60% and 63% for strains M8 and M16 against B. cinerea and between 40% and 53% for strains M8 and M16 against F. solani. Based on these results, the need to implement resistance induction against gray mold on pepper plants was determined using strains M8 and M16. In this case, strain M16 inhibited the propagation of the necrotic spot by approximately 70%, whereas strain M8 significantly reduced the superoxide dismutase activity in systemic leaves, which substantially increased in plants inoculated with strain M8 and infected with the pathogen. Accordingly, the use of native rhizobacteria may entail biotechnological progress for the integrated management of crops in agriculture industry.

14.
Biotechnol Rep (Amst) ; 26: e00461, 2020 Jun.
Artigo em Inglês | MEDLINE | ID: mdl-32420051

RESUMO

Coinoculation of plants with mixtures of beneficial microbes sometimes produces synergistic effects. In this study, the effect of soybean coinoculation with the N2-fixing Bradyrhizobium japonicum E109 and the biocontrol fungus Trichoderma harzianum Th5cc was analyzed. Nodulation by E109 was not hampered by Th5cc, which antagonized five out of seven soybean pathogens tested. Furthermore, Th5cc relieved nitrate-inhibition of nodulation, enabling the formation of nodules containing infected cells with bacteroids in the presence of the otherwise inhibitory 10 mM KNO3. Th5cc released micromolar amounts of auxin, and addition of 11 µM indoleacetic acid to soybean plants inoculated with E109 in the absence of Th5cc also induced nodulation in the presence of 10 mM KNO3. Thus, Th5cc may release auxins into the soybean rhizosphere, which hormones might participate in overcoming the nitrate-inhibition of nodulation. Our results suggest that soybean plants coinoculated with these microorganisms might benefit from biocontrol while contributing to soil-nitrogen preservation.

15.
Arch Microbiol ; 201(2): 171-183, 2019 Mar.
Artigo em Inglês | MEDLINE | ID: mdl-30535938

RESUMO

Rhizobium tropici CIAT 899 is a strain known by its ability to nodulate a broad range of legume species, to synthesize a variety of Nod factors, its tolerance of abiotic stresses, and its high capacity to fix atmospheric N2, especially in symbiosis with common bean (Phaseolus vulgaris L.). Genes putatively related to the synthesis of indole acetic acid (IAA) have been found in the symbiotic plasmid of CIAT 899, in the vicinity of the regulatory nodulation gene nodD5, and, in this study, we obtained mutants for two of these genes, y4wF and tidC (R. tropiciindole-3-pyruvic acid decarboxylase), and investigated their expression in the absence and presence of tryptophan (TRP) and apigenin (API). In general, mutations of both genes increased exopolysaccharide (EPS) synthesis and did not affect swimming or surface motility; mutations also delayed nodule formation, but increased competitiveness. We found that the indole-3-acetamide (IAM) pathway was active in CIAT 899 and not affected by the mutations, and-noteworthy-that API was required to activate the tryptamine (TAM) and the indol-3-pyruvic acid (IPyA) pathways in all strains, particularly in the mutants. High up-regulation of y4wF and tidC genes was observed in both the wild-type and the mutant strains in the presence of API. The results obtained revealed an intriguing relationship between IAA metabolism and nod-gene-inducing activity in R. tropici CIAT 899. We discuss the IAA pathways, and, based on our results, we attribute functions to the y4wF and tidC genes of R. tropici.


Assuntos
Carboxiliases/metabolismo , Ácidos Indolacéticos/metabolismo , Rhizobium tropici/genética , Rhizobium tropici/metabolismo , Carboxiliases/genética , Genes Bacterianos , Indóis/metabolismo , Mutação , Phaseolus/microbiologia , Phaseolus/fisiologia , Polissacarídeos Bacterianos/biossíntese , Rhizobium tropici/química , Rhizobium tropici/enzimologia , Simbiose
16.
Front Microbiol ; 9: 1907, 2018.
Artigo em Inglês | MEDLINE | ID: mdl-30186252

RESUMO

In recent years, Chilean kiwifruit production has been affected by the phytopathogen Pseudomonas syringae pv. actinidiae (Psa), which has caused losses to the industry. In this study, we report the genotypic and phenotypic characterization of 18 Psa isolates obtained from Chilean kiwifruits orchards between 2012 and 2016 from different geographic origins. Genetic analysis by multilocus sequence analysis (MLSA) using four housekeeping genes (gyrB, rpoD, gltA, and gapA) and the identification of type III effector genes suggest that the Chilean Psa isolates belong to the Psa Biovar 3 cluster. All of the isolates were highly homogenous in regard to their phenotypic characteristics. None of the isolates were able to form biofilms over solid plastic surfaces. However, all of the isolates formed cellular aggregates in the air-liquid interface. All of the isolates, except for Psa 889, demonstrated swimming motility, while only isolate Psa 510 demonstrated swarming motility. The biochemical profiles of the isolates revealed differences in 22% of the tests in at least one Psa isolate when analyzed with the BIOLOG system. Interestingly, all of the isolates were able to produce indole using a tryptophan-dependent pathway. PCR analysis revealed the presence of the genes aldA/aldB and iaaL/matE, which are associated with the production of indole-3-acetic acid (IAA) and indole-3-acetyl-3-L-lysine (IAA-Lys), respectively, in P. syringae. In addition, IAA was detected in the cell free supernatant of a representative Chilean Psa strain. This work represents the most extensive analysis in terms of the time and geographic origin of Chilean Psa isolates. To our knowledge, this is the first report of Psa being able to produce IAA. Further studies are needed to determine the potential role of IAA in the virulence of Psa during kiwifruit infections and whether this feature is observed in other Psa biovars.

17.
Proc Natl Acad Sci U S A ; 115(21): 5612-5617, 2018 05 22.
Artigo em Inglês | MEDLINE | ID: mdl-29724856

RESUMO

Light cues from neighboring vegetation rapidly initiate plant shade-avoidance responses. Despite our detailed knowledge of the early steps of this response, the molecular events under prolonged shade are largely unclear. Here we show that persistent neighbor cues reinforce growth responses in addition to promoting auxin-responsive gene expression in Arabidopsis and soybean. However, while the elevation of auxin levels is well established as an early event, in Arabidopsis, the response to prolonged shade occurs when auxin levels have declined to the prestimulation values. Remarkably, the sustained low activity of phytochrome B under prolonged shade led to (i) decreased levels of PHYTOCHROME INTERACTING FACTOR 4 (PIF4) in the cotyledons (the organs that supply auxin) along with increased levels in the vascular tissues of the stem, (ii) elevated expression of the PIF4 targets INDOLE-3-ACETIC ACID 19 (IAA19) and IAA29, which in turn reduced the expression of the growth-repressive IAA17 regulator, (iii) reduced abundance of AUXIN RESPONSE FACTOR 6, (iv) reduced expression of MIR393 and increased abundance of its targets, the auxin receptors, and (v) elevated auxin signaling as indicated by molecular markers. Mathematical and genetic analyses support the physiological role of this system-level rearrangement. We propose that prolonged shade rewires the connectivity between light and auxin signaling to sustain shade avoidance without enhanced auxin levels.


Assuntos
Proteínas de Arabidopsis/metabolismo , Arabidopsis/metabolismo , Regulação da Expressão Gênica de Plantas/efeitos dos fármacos , Ácidos Indolacéticos/farmacologia , Luz , Fitocromo/metabolismo , Fenômenos Fisiológicos Vegetais , Arabidopsis/efeitos dos fármacos , Arabidopsis/crescimento & desenvolvimento , Proteínas de Arabidopsis/genética , Fitocromo/genética , Reguladores de Crescimento de Plantas/farmacologia , Transdução de Sinais
18.
Plant Physiol Biochem ; 127: 506-515, 2018 Jun.
Artigo em Inglês | MEDLINE | ID: mdl-29709880

RESUMO

In semi-arid regions, plants develop various biochemical and physiological strategies to adapt to dry periods. Understanding the resistance mechanisms to dry periods under field conditions is an important topic in ecology. Larrea divaricata and Lycium chilense provide various ecological services. The aim of this work is to elucidate new morpho-histological, biochemical and hormonal traits that contribute to the drought resistance strategies of two native shrubs. Green leaves and fine roots from L. divaricata and L. chilense were collected in each season for one year, and various traits were measured. The hormone (abscisic acid, ABA-glucose ester, gibberellins A1 and A3, and indole acetic acid) contents were determined by liquid chromatography coupled to mass spectrometry. Rainfall data and the soil water content were also measured. A multivariate analysis showed that green leaves from L. divaricata showed high values for the leaf dry weight, blade leaf thickness and ABA content in the summer compared with those from L. chilense. Fine roots from L. divaricata had high RWC and high IAA levels during the autumn-dry period compared with those from L. chilense, but both had similar levels during the winter and spring. Our results support the notion that species with different drought resistance mechanisms (avoidance or tolerance) display different responses to dry periods throughout the year. Larrea divaricata, which exhibits more xerophytic traits, modified its morphology and maintained its physiological parameters (high RWC in leaves and roots, high ABA levels in leaves during summer, high GA3 in leaves and high IAA in roots during autumn) to tolerate dry periods, whereas Lycium chilense, which displays more mesophytic traits, uses strategies to avoid dry periods (loss of leaves during autumn and winter, high RWC in leaves, high ABA-GE and GA3 in leaves during summer, high GA1 and GA3 in roots during summer, and high IAA in roots during autumn and summer) and thus has a metabolism that is more dependent on water availability for growth.


Assuntos
Larrea/crescimento & desenvolvimento , Lycium/crescimento & desenvolvimento , Folhas de Planta/crescimento & desenvolvimento , Raízes de Plantas/crescimento & desenvolvimento , Estações do Ano , Argentina , Desidratação/metabolismo , Especificidade da Espécie
19.
Braz. J. Microbiol. ; 49(1): 59-66, jan.-mar. 2018. ilus, tab
Artigo em Inglês | VETINDEX | ID: vti-19065

RESUMO

Mangroves are ecosystems located in the transition zone between land and sea that serve as a potential source of biotechnological resources. Brazil's extensive coast contains one of the largest mangrove forests in the world (encompassing an area of 25,000 km2 along all the coast). Endophytic bacteria were isolated from the following three plant species: Rhizophora mangle, Laguncularia racemosa and Avicennia nitida. A large number of these isolates, 115 in total, were evaluated for their ability to fix nitrogen and solubilize phosphorous. Bacteria that tested positive for both of these tests were examined further to determine their level of indole acetic acid production. Two strains with high indole acetic acid production were selected for use as inoculants for reforestation trees, and then the growth of the plants was evaluated under field conditions. The bacterium Pseudomonas fluorescens (strain MCR1.10) had a low phosphorus solubilization index, while this index was higher in the other strain used, Enterobacter sp. (strain MCR1.48). We used the reforestation tree Acacia polyphylla. The results indicate that inoculation with the MCR1.48 endophyte increases Acacia polyphylla shoot dry mass, demonstrating that this strain effectively promotes the plant's growth and fitness, which can be used in the seedling production of this tree. Therefore, we successfully screened the biotechnological potential of endophyte isolates from mangrove, with a focus on plant growth promotion, and selected a strain able to provide limited nutrients and hormones for in plant growth.(AU)


Assuntos
Acacia/crescimento & desenvolvimento , Acacia/microbiologia , Rhizophoraceae , Endófitos , Áreas Alagadas , Avicennia , Combretaceae
20.
Braz. j. microbiol ; Braz. j. microbiol;49(1): 59-66, Jan.-Mar. 2018. tab, graf
Artigo em Inglês | LILACS | ID: biblio-889203

RESUMO

ABSTRACT Mangroves are ecosystems located in the transition zone between land and sea that serve as a potential source of biotechnological resources. Brazil's extensive coast contains one of the largest mangrove forests in the world (encompassing an area of 25,000 km2 along all the coast). Endophytic bacteria were isolated from the following three plant species: Rhizophora mangle, Laguncularia racemosa and Avicennia nitida. A large number of these isolates, 115 in total, were evaluated for their ability to fix nitrogen and solubilize phosphorous. Bacteria that tested positive for both of these tests were examined further to determine their level of indole acetic acid production. Two strains with high indole acetic acid production were selected for use as inoculants for reforestation trees, and then the growth of the plants was evaluated under field conditions. The bacterium Pseudomonas fluorescens (strain MCR1.10) had a low phosphorus solubilization index, while this index was higher in the other strain used, Enterobacter sp. (strain MCR1.48). We used the reforestation tree Acacia polyphylla. The results indicate that inoculation with the MCR1.48 endophyte increases Acacia polyphylla shoot dry mass, demonstrating that this strain effectively promotes the plant's growth and fitness, which can be used in the seedling production of this tree. Therefore, we successfully screened the biotechnological potential of endophyte isolates from mangrove, with a focus on plant growth promotion, and selected a strain able to provide limited nutrients and hormones for in plant growth.


Assuntos
Bactérias/isolamento & purificação , Árvores/microbiologia , Acacia/microbiologia , Endófitos/isolamento & purificação , Filogenia , Bactérias/classificação , Bactérias/genética , Bactérias/metabolismo , Árvores/crescimento & desenvolvimento , Brasil , Acacia/crescimento & desenvolvimento , Áreas Alagadas , Endófitos/classificação , Endófitos/genética , Endófitos/metabolismo , Ácidos Indolacéticos/metabolismo
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